Anti-allergy composition and related method

ABSTRACT

An composition and related method that prevents, inhibits and/ or mitigates an allergic response by down regulating the production of IgE, down regulating the binding of IgE antibodies to receptors on cells, and/or inhibiting allergy mediators, for example, histamine, prostoglandin D 2 , or luekotriene C 4  release. The composition comprises at least one of the following ingredients: luteolin from Perilla leaf or seed, Cinnamon, Kiwi, Picao preto, Hesperidin, Acerola cherry, Guaco, Holy Basil, Kakadu, Solamum, Rosmarinic acid, Tinospora and Aframomum. In one embodiment, the composition at least three different ingredients selected from Cinnamon, Acerola, Luteolin and Picao preto. Optionally, these ingredients can be combined with at least one of Aframomum, Rosmarinic acid, and Tinospora. The composition is administered with effective amounts to prevent, inhibit and/or mitigate allergic responses.

BACKGROUND OF THE INVENTION

This application claims benefit of U.S. Provisional Application No.60/542,070, filed Feb. 5, 2004, which is hereby incorporated byreference.

The present invention relates to a composition and method for preventingand treating allergic reactions and diseases.

Many individuals are affected by allergy-related immunologicaldisorders. Such disorders include, for example, bronchial asthma,allergic rhinitis (hay fever), and atopic dermatitis. Although thesedisorders have somewhat different effects on the body, they share acommon allergic response, referred to as a type I, or immediate-type,anaphylactic allergic reaction. This response is mediated by substances,such as immunoglobin E (IgE) and histamines, and includes several steps.

In the first step of the reaction, referred to as the sensitizationstep, an immune response to an allergen is initiated upon exposure of aresponsive individual to the allergen. This results in the generation ofB cells that secrete allergen specific IgE. The IgE subsequently bindsto IgE receptor sites on mast cells and basophils. In a second step,referred to as the degranulation step, when re-exposed to the allergen,the aforementioned receptor-bound IgE binds to the allergen resulting indegranulation of the mast cells and basophils. Degranulation releases avariety of vasoactive mediators, for example, histamines and proteases,which subsequently promote allergic and inflammatory responses. Thethird step, referred to as the inflammation step, is triggered by themediators, and causes inflammatory cells to a) accumulate at sites ofinflammation, for example, target organs, such as the lungs, and b)release chemicals, such as interleukin-3, interleukin-6, and macrophagecolony stimulating factors. While inflammatory cells are normallyactivated to provide tissue defense, tissue maintenance andimmunoregulation, in the case of allergies, activation of inflammatorycells serves to augment the allergic response.

Several allergic disease treatments exist. Many of these treatments usesteroids to either inhibit the release of substances during thedegranulation step, or inhibit the allergic reactions induced during theinflammation step. Although effective in mitigating allergic responses,these treatments are unhelpful in preventing the onset of the type Iallergic reaction, that is, the treatments fail to inhibit IgEproduction, and accordingly fail to stop the underlying inflammatoryprocess. A consequence of this is that the treatments also fail toprevent an often life-threatening reaction referred to as anaphylacticshock, which is triggered simply by the initiation of the type Iallergic reaction. Further, many of drugs used to inhibit thedegranulation and inflammation steps also have undesirable side effects.For example, degranulation is often treated with corticosteroids, andinflammation is frequently treated with glucocortosteroids; however,these drugs can cause numerous side effects, such as weight gain, waterretention, hypertension, and increased cholesterol levels.

SUMMARY OF THE INVENTION

The aforementioned problems are overcome in the present invention whichprovides a composition and related method that inhibits or prevents anallergic response by down regulating the production of IgE, downregulating the binding of IgE antibodies to receptors on cells, and/orinhibiting histamine, prostoglandin D₂, or luekotriene C₄ release.

In one embodiment, the composition and method reduce and/or prevent theproduction of IgE antibodies. This can be accomplished by thecomposition preventing B cells of the responsive individual fromsecreting IgE antibodies.

In another embodiment, the composition and method reduces the level ofIgE receptor sites on mast cells and basophils. This, in turn, canprevent the activation of the mast cells and/or basophils to releasethose mediator substances that promote allergic and subsequentinflammatory responses.

In a further embodiment, the composition and method inhibits or preventsthe release of allergic response mediators, including histamine,prostoglandin D₂, and/or luekotriene C₄. Accordingly, cell degranulationcan be prevented to prevent, inhibit or stop an allergic or inflammatoryresponse.

In yet another embodiment, the composition can include at least one ofthe following ingredients: Perilla leaf (or Perilla seed), specificallythe ingredient luteolin (from Perilla fruteucens), Cinnamon (Cinnamonzeylanicum), Kiwi extract (from Actinidia arguta), Picao preto (Bidenspilosa), Hesperidin (Citrus senesis), Acerola cherry (Malphighiaglabra), Guaco (Mikania glomerata), Holy Basil (Ocimum sanctum), Kakadu(Terminalia fernandida), Solarnum (Solanum xanthocarpum), Kiwi juice(from Actinidia chinensis), Rosmarinic acid (from Rosmarinusofficinalis), Tinospora (from Tinospora cordifolia), and Aframomum (fromAframomum melegueta). In a more specific embodiment, the composition caninclude: Cinnamon, Acerola and Picao preto. In an even more specificembodiment, the composition can include: Cinnamon, Acerola, Picao pretoand at least one of luteolin, Tinospora, Rosmarinic acid, and Aframomum.These ingredients can be present in equal amounts. In another morespecific embodiment, the composition can include: Cinnamon, Acerola,luteolin and at least one of Picao preto, Tinospora, Rosmarinic acid,and Aframomum. These ingredients can be present in equal amounts.

In a method of the present invention, the compositions above can beadministered to cells and/or subjects to inhibit or prevent an allergicor inflammatory response by at least one of (a) down regulating theproduction of IgE, (b) down regulating the binding of IgE antibodies toreceptors on cells, and/or (c) inhibiting or preventing the release ofmediators such as histamine, PGD₂ and/or LTC₄.

In the method of the present invention, the compositions above can beadministered in an effective amount to a subject undergoing an allergicresponse or disease or the potential for the same, for example, allergicrhinitis, bronchial asthma, allergic conjunctivitis, atopic dermatitis,food allergy, hyper IgE syndrome, anaphylactic shock, atopic eczema andrheumatoid arthritis. In one embodiment, the composition can beadministered to lower IgE levels in a subject, for example, by downregulating the production of and/or binding of IgE antibodies toreceptors on cells. In another embodiment, the composition can beadministered to inhibit or prevent the release of allergic responsemediators such as histamine, PGD₂ and/or LTC₄.

The present invention provides many benefits over many conventionalanti-allergy treatments. For example, the composition and method caninhibit IgE production, and accordingly prevent or delay sensitizationof mast cells and/or basophils, and the subsequent inflammatory process.Additionally, the use of ingredients optionally derived from fruit andherb sources in the composition can reduce the potential for adverseside effects for subjects using the composition. Further, thecomposition can target both allergic responses upstream of most currentanti-allergy treatments, which only block histamine action or preventthe production of anti-inflammatory mediators such as prostoglandins andleukotrienes, as well as inhibit or prevent release of theanti-inflammatory mediators. Accordingly, the present invention canmitigate and/or prevent the incidence of anaphylactic shock, which manyother purely downstream treatments fail to do, as well as down regulateallergic response mediators.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph illustrating the effect of the compositions of thepresent invention on IgE receptor expression;

FIG. 2 is a graph illustrating the expression of different IgE receptorson test cells;

FIG. 3 is a graph illustrating the effect of the compositions on IgEsecretion;

FIG. 4 is a graph illustrating a calibration curve used to extrapolateIgE concentration from the optical density data of composition samples;

FIG. 5 is a graph illustrating the effect of individual ingredientcompositions at varying concentrations on IgE receptor expression;

FIG. 6 is a graph illustrating the effect of individual ingredientcompositions at varying concentrations on IgE secretion;

FIG. 7 is a graph illustrating the effect of the ingredient compositionson IgE receptor expression;

FIG. 8 is a graph illustrating the effect of the ingredient compositionson IgE secretion;

FIG. 9 is a graph illustrating the effect of the ingredient compositionson allergy mediator release;

FIG. 10 is a graph illustrating a calibration curve used to extrapolatehistamine release from the optical density data of composition samples;

FIGS. 11-13 are graphs illustrating the effect of a multiple ingredientcompositions on mediator release; and

FIG. 14 is a graph illustrating the effect of multiple ingredientcompositions on IgE secretion, IgE-Receptor expression and histaminerelease.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

I. Composition and Manufacture

The composition of the present invention can be prepared by mixingacceptable carriers with a mixture including effective amounts of atleast one, at least two, at least three or at least four of thefollowing ingredients: Perilla leaf or seed, specifically the ingredientluteolin (from Perilla fruteucens), Cinnamon (Cinnamon zeylanicum), Kiwiextract (from Actinidia arguta), Picao preto (Bidens pilosa), Hesperidin(Citrus senesis), Acerola cherry (Malphighia glabra), Guaco (Mikaniaglomerata), Holy Basil (Ocimum sanctum), Kakadu (Terminalia fernandida),Solarnum (Solanum xanthocarpum), Kiwi juice (from Actinidia chinensis),Rosmarinic acid (from Rosmarinus officinalis), Tinospora (from Tinosporacordifolia), Aframomum (from Aframomum melegueta), Tumeric, (curcumalonga), Cumaru (Amburana cearensis), Marigold (Tagetes erecta), NettleRoot (Urtica dioica), Quercetin (Dimorphandra gardneriana), Ginger(Zingiber officinale), Boswin 30 (Borwellia serratta), Trikartu extract(An ayurvedic extract composed of a third each of Pipli (piper longum),Black pepper (piper nigrum), Bioperine, available from Sabinsa Corp. ofPiscataway, N.J., and Allergy Formula, available from Geni Herbs ofNoblesville, Ind.

In one embodiment, the composition includes: Cinnamon, Acerola and Picaopreto. Optionally, this composition further includes at least one ofluteolin, Tinospora, Rosmarinic acid, and Aframomum. In anotherembodiment, the composition includes: Cinnamon, Acerola and luteolin.Optionally, this composition further includes at least one of Picaopreto, Tinospora, Rosmarinic acid, and Aframomum. When multipleingredients are included in the composition, the ingredients can bepresent in approximately equal amounts by weight of the composition. Forexample, a composition can include 10% by weight carriers, and 90% byweight IgE and/or mediator affecting ingredients. Where there are threeingredients in this exemplary composition, such as Cinnamon, Acerola andPicao preto, each of these ingredients may be present in amounts eachequal to about 30% by weight of the composition.

Some ingredients of the composition perform better at affecting orotherwise inhibiting certain mechanisms in allergic and inflammatoryresponses. For example, some ingredients down regulate IgE via aspecific mechanism. As a specific example, Kiwi extract operatesextremely well to reduce IgE secretion, but moderately well at reducingIgE receptor expression. On the other hand, Cinnamon operates very wellto reduce IgE receptor expression, but moderately well to reduce IgEsecretion. Accordingly, some compositions of the present inventioninclude both ingredients that reduce IgE receptor expression, andingredients that reduce IgE secretion. These compositions therefore downregulate IgE via two mechanisms: by reducing IgE receptor expression andby reducing IgE secretion. As used herein, allergic response encompassesan inflammatory response, and inhibiting an allergic response includes,but is not limited to partial inhibition of the response, completearrest of the response, and/or prevention of the response.

Furthermore, some ingredients additionally or alternatively inhibit therelease of mediators better than other ingredients; and some ingredientsinhibit the release of some mediators better than other mediators. As aspecific example, Aframomum operates better than Cinnamon at inhibitingLTC₄ release, but Cinnamon outperforms Aframomum at inhibition ofhistamine release. Likewise, Aframomum operates very well to inhibitLTC₄ release, but only moderately well to inhibit histamine release.Moreover, some ingredients, for example Cinnamon, affect severalmechanisms in the allergic response. As noted above, Cinnamon reducesIgE receptor expression, as well as inhibits the release of mediatorssuch as histamine and PGD₂. The specific mechanisms by which thedifferent ingredients and combinations of ingredients operate, as wellas their relative efficacy, is discussed in further detail below.

Compositions that included specific combinations of ingredients alsoexhibited surprising and unexpected synergy in affecting certainmechanisms. For example, the combination of Cinnamon, Acerola and Picaopreto, also referred to herein as the Blend, inhibited histamine releasebetter than any one of these ingredients alone. As discussed in furtherdetail below in Example 7, the Blend inhibited histamine release to 29%of the histamine release by cells untreated with the Blend or any otheringredient. In contrast, Cinnamon individually inhibited histaminerelease to 84%, Acerola to 70% and Picao preto to 96%. This result wasinteresting because a combination of ingredients does not frequentlylead to such synergy and overall improvement in mechanism effect. Forexample, in Example 8, the Blend inhibited histamine release to 73% ofthat by untreated control cells, Aframomum inhibited histamine releaseto 96% of untreated control samples, but the combination of the Blendand Aframomum inhibited histamine release to 100% of untreated controlsamples. It is believed that the frequent non-synergistic effect ofingredient combinations is caused by the individual ingredientscounteracting one another, and/or affecting different biologicalmechanisms, so that the net effect of the combination is less than, oris less desirable than, the effect of each ingredient alone.

For purposes of this disclosure, an effective amount of an ingredient orcomposition refers to the amount necessary to elicit the desiredbiological response. Additionally, as used herein, mast cell refers toat least one of mast cells, basophils, and other cells with IgEreceptors. Further, the Ginger can be of a special variety, for exampleGinger 5%, which means Ginger extract at 5% concentration. In oneembodiment, the Kiwi extract used is of a special variety, referred toas PanGenomic A, which is available from PanGenomic Co., Ltd. of Seoul,Korea. The Hesperidin used can also be of a special variety,specifically Hesperidin 80%, which means Hesperidin extract at 80%concentration. In addition, the Tinospora used can be Tinospora presentin the product sold under the trade name Tinofend, which is commerciallyavailable from Geni Herbs.

Effective amounts for particular embodiments of the composition whenused in connection with in-vitro testing may vary as desired. In oneembodiment, the composition includes a combination of the ingredientsCinnamon, Acerola and Picao preto, referred to herein as a Blend, withranges of dosages for the Blend being greater than 1 μg/ml, with eachingredient of the Blend being present in equal concentrations.

In another embodiment, at least one of Luteolin, Aframomum, Rosmarinicacid, and Tinospora is added to the Blend. The Luteolin may be derivedfrom Perilla seed and/or optionally, Perilla leaf. In this embodiment,the dosage range for the Luteolin may be greater than 1 μg/ml, thedosage range for the Aframomum may be greater than 1 μg/ml, the dosagerange for the Rosmarinic acid may be greater than 1 μg/ml, and thedosage range for the Tinospora may be greater than 1 μg/ml.

Furthermore, effective amounts for particular embodiments of thecomposition when used in connection with in-vivo testing or regularsubject administration may vary as desired. In one embodiment, thecomposition may include a combination of the ingredients Cinnamon,Acerola and Picao preto, i.e., the Blend, with ranges of daily subjectdosages for the Blend being from about 1 mg to about 540 mg; about 1 mgto about 400 mg; about 1 mg to about 270 mg; about 270 mg to about 400mg; or about 270 mg to about 540 mg. In this embodiment, each individualingredient of the Blend may have ranges of daily subject dosages thatare equal to that of the Blend ranges. The components of the Blend alsocan be expressed as a percent by weight of the total composition inwhich the Blend is incorporated. Specifically, Cinnamon can be presentin a percent by weight in the ranges of: about 5% to about 50%; about10% to about 25%; about 15% to about 22%; and about 20%. Acerola may bepresent in the amount by weight percent of: about 5% to about 50%; about10% to about 25%; about 15% to about 22%; and about 20%. Picao preto maybe present in a percent by weight in the ranges of: about 5% to about50%; about 10% to about 25%; about 15% to about 22%; and about 20%.Other ranges may be used as desired.

In a more specific embodiment, at least one of Luteolin, Aframomum,Rosmarinic acid, and Tinospora can be added to the Blend. The Luteolinmay be derived from Perilla seed and/or optionally, Perilla leaf. Inthis embodiment, the ranges of daily subject dosages for the Luteolinmay be from about 1 mg to about 225 mg; about 1 mg to about 180 mg;about 1 mg to about 135 mg; about 135 mg to about 180 mg; or about 180mg to about 225 mg. The ranges of daily subject dosages in thisembodiment for the Aframomum or Rosmarinic acid may be from about 1 mgto about 300 mg; about 1 mg to about 225 mg; about 1 mg to about 150 mg;about 150 mg to about 225 mg; or about 225 mg to about 300 mg. Theranges of daily subject dosages in this embodiment for the Tinospora maybe from about 1 mg to about 900 mg; about 1 mg to about 600 mg; about 1mg to about 300 mg; about 300 mg to about 600 mg; or about 600 mg toabout 900 mg.

In yet another embodiment, the composition may include a combination ofthe ingredient Cinnamon, Acerola and Luteolin, a combination which isalso referred to as the Formula herein. The Luteolin may be derived fromPerilla seed and/or optionally, Perilla leaf. Acceptable effectiveamounts for this embodiment when used in connection with in-vitrotesting may vary as desired. An example of such a dosage range for theFormula is greater than 1 μg/ml, with each ingredient of the Formulabeing present in equal concentrations.

In another embodiment, at least one of Picao preto, Luteolin, Aframomum,Rosmarinic acid, and Tinospora is added to the Formula. In thisembodiment, the dosage range for the Picao preto may be greater than 1μg/ml, the dosage range for the Aframomum may be greater than 1 μg/ml,the dosage range for the Rosmarinic acid may be greater than 1 μg/ml,and the dosage range for the Tinospora may be greater than 1 μg/ml.

Acceptable ranges of daily subject dosages for the Formula are fromabout 1 mg to about 540 mg; about 1 mg to about 400 mg; about 1 mg toabout 270 mg; about 270 mg to about 400 mg; or about 270 mg to about 540mg. In this embodiment, each individual ingredient of the Formula mayhave ranges of daily subject dosages that are equal to that of theFormula ranges. In another example, the components of the Formula can beexpressed as a percent by weight of the total composition in which theFormula is incorporated. In such an example, Cinnamon can be present ina percent by weight in the ranges of: about 5% to about 50%; about 10%to about 25%; about 15% to about 22%; and about 20%. Acerola may bepresent in the amount by weight percent of: about 5% to about 50%; about10% to about 25%; about 15% to about 22%; and about 20%. Luteolin may bepresent in a percent by weight in the ranges of: about 5% to about 50%;about 10% to about 25%; about 15% to about 22%; and about 20%. Otherranges may be used as desired. In a more specific embodiment, at leastone of Picao preto, Aframomum and Tinospora may be added to the Formula.The ranges of daily subject dosages for these ingredients may the sameas that noted above in connection with the previous embodiment.

The composition of the present invention can be prepared in any mannerthat preserves the biological activity of the ingredients. Possiblepreparations of the composition include decoctions, aqueous extracts,organic solvent extracts, and dry powder. In one embodiment, theingredients are dried and ground, and the resulting powder is processedinto pill form, however, the composition can be processed into formshaving varying delivery systems. For example, the ingredients can beprocessed and included in capsules, tablets, gel tabs, lozenges, strips,granules, powders, concentrates, solutions or suspensions. Theingredients can also be administered into the respiratory tract, e.g. inthe treatment of asthma, anaphylactic or and other acute shockconditions via a spray, mist or aerosol. The ingredients also can beformulated, individually or in combination, with other foods to providepre-measured nutritional supplements, supplemented foods, for example,single serving bars. In general, the composition can be administered tosubjects orally, parenterally, rectally, intracisternally,intraperitoneally, topically and bucally.

The compositions of the present invention can include at least oneacceptable excipient or carrier. For purposes of this disclosure,acceptable carrier means a non-toxic, inert solid, semi-solid or liquidfiller, diluent, encapsulating material or formulation. Examples ofacceptable carriers are the following: cellulose and its derivatives,such as sodium carboxymethyl cellulose, ethyl cellulose and celluloseacetate; starches, such as corn and potato starches; sugars, such aslactose, glucose, and sucrose; gelatin; talc; excipients, such as cocoabutter and waxes; oils, such as peanut oil, cottonseed oil, saffloweroil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such aspropylene glycol; esters, such as ethyl oleate and ethyl laurate; agar;buffering agents, such as magnesium hydroxide and aluminum hydroxide;alginic acid; pyrogen-free water; isotonic saline; Ringer's solution;ethyl alcohol; and phosphate buffer solutions. Other agents that can bepresent in the composition of the present invention include sweeteningand flavoring agents, coating agents, lubricants, flow aids, coloringagents, releasing agents, perfuming agents, preservatives andantioxidants, as the application requires.

As a specific example of a delivery vehicle, preselected amounts ofmultiple ingredients, e.g., Cinnamon, Acerola and Picao preto in any ofthe ranges presented above, can be blended with excipients includingmicrocrystalline cellulose silicified (a filler), stearic acid (alubricant), and silicon dioxide (a flow aid). The blend can be filledinto size #0 white capsules using a hand operated capsule fillingmachine, commercially available from T.M.P. of Milano, Italy. Suchcapsules can be administered to subjects according to a dosage regimenas desired.

II. Method of Use

The compositions of the present invention can be administered in avariety of ways. Specifically, the compositions can be administered tocells and/or a subject alone, or in combination with additionaltherapeutic treatments, such as corticosteroids, which are acceptabledrugs for treating asthma. Moreover, the compositions can beadministered with standard or reduced corticosteroid or othertreatments. Further, the compositions can be administered in combinationwith other acceptable drugs use to treat allergies including, but notlimited to, anti-histamines, non-steroidal anti-inflammatory drugs,steroidal anti-inflammatory drugs and decongestants.

In one embodiment, the compositions can be employed to treat existingallergic responses. For example, the compositions can be used to reducethe severity, intensity, and/or duration of an asthmatic response. Insuch an application, the compositions are administered to an individualafter the individual develops the asthmatic symptoms. The compositionscan also be administered to cells to test effect of the composition onan allergic response of the cells.

In an alternative or additional embodiment, the compositions can be usedto prevent or delay the onset of a type I allergic reaction in anindividual who has previously suffered such a reaction, or to prevent,reduce the severity, intensity, or duration of subsequently-developedsymptoms. Optionally, where an antigen has been identified that is knownto have induced, or at least is correlated with, the onset of previousallergic reactions, the compositions can be administered either beforethe onset of symptoms after a subsequent encounter with the antigen, orbefore the encounter.

In another alternative or additional embodiment, the compositions can beadministered before the development of allergic sensitivity to aparticular antigen. In this case, the compositions can be administeredsubstantially concurrently with exposure to an antigen that has notpreviously been associated with an allergic reaction.

In general, the compositions of the present invention can beadministered in an effective amount to cells or a subject having anallergic response or disease or the potential for the same, for example,allergic rhinitis, bronchial asthma, allergic conjunctivitis, atopicdermatitis, food allergy, hyper IgE syndrome, anaphylactic shock, atopiceczema and rheumatoid arthritis.

In one embodiment, the composition can be administered to inhibit anallergic response. This can be done by lowering IgE levels in cells or asubject, for example, by down regulating the production of and/orbinding of IgE antibodies to receptors on cells. In turn, this isbelieved to prevent, mitigate and/or inhibit the sensitization step of atype I allergic reaction. By preventing, mitigating and/or inhibiting(all of which are encompassed by the term inhibiting as used herein) theonset of a type I allergic reaction, the subsequent reaction andresulting symptoms can also be prevented, mitigated and/or inhibited.These symptoms include but are not limited to hives, rashes, puritis,watery eyes, edema, diarrhea, bronchial constriction and/orinflammation, airway hyper-responsiveness, difficulty breathing,vasodilation, a decrease in blood pressure, increased IgE levels,increased plasma histamine levels, increased protease levels,anaphylaxis, and death.

In a further embodiment, the composition can be administered to cells ora subject to inhibit or prevent the release of allergic reactionmediators such as histamine, PGD₂ and/or LTC₄. In so doing, the type Iallergic reaction, the subsequent reaction and resulting symptoms canalso be inhibited.

III. Identification and Characterization of Compositions

A. IgE Testing and Results

The effects of the composition on IgE can be analyzed in any in-vivo orin-vitro model. In an exemplary in-vitro model, such testing wasconducted by treating a basophilic cell line having cells with highlevels of IgE receptors on their surfaces. After a predefined incubationperiod, the level of IgE secretion and IgE receptor expression wasmeasured with immunochemical methods. The following examples describehow IgE secretion and IgE receptor expression was measured to identifyand select combinations of ingredients having acceptable IgEdown-regulating effect. Although Examples 1 and 3 describe ways toidentify combinations of ingredients having IgE down-regulating effects,the methodologies set forth can also or alternatively be used toidentify individual ingredients having IgE down-regulating effects, asillustrated in Examples 2, 4 and 5.

It is noted that the following examples are intended to illustratecertain particular embodiments of the invention, but are not intended tolimit their scope, as defined by the claims.

EXAMPLE 1

Multiple combinations of composition ingredients, presented in Table I,were tested to determine the each of the combination's effectiveness incontrolling IgE receptor expression. In each of the compositions ofTable I, the ingredients are present in equal amounts. FIG. 1demonstrates the effects of the compositions on IgE receptor expression.TABLE I Sample Composition Combinations of Ingredients 5 ingredients 4ingredients 3 ingredients Perilla seed 4-1 Perilla seed 3-1 Perilla seedHesperidin 80% Hesperidin 80% Hesperidin 80% Picao preto Picao pretoPicao preto Kiwi extract Kiwi extract 3-2 Perilla seed Cinnamomum 4-2Perilla seed Hesperidin 80% Hesperidin 80% Kiwi extract Picao preto 3-3Perilla seed Cinnamomum Hesperidin 80% 4-3 Perilla seed CinnamomumHesperidin 80% 3-4 Perilla seed Kiwi extract Picao preto Cinnamomum Kiwiextract 4-4 Perilla seed 3-5 Perilla seed Picao preto Picao preto Kiwiextract Cinnamomum Cinnamomum 3-6 Perilla seed 4-5 Hesperidin 80% Kiwiextract Picao preto Cinnamomum Kiwi extract 3-7 Hesperidin 80%Cinnamomum Picao preto Cinnamomum 3-8 Picao preto Kiwi extractCinnamomum 3-9 Hesperidin 80% Kiwi extract Cinnamomum  3-10 Hesperidin80% Picao preto Kiwi extract

Specifically, FIG. 1 presents the combination of ingredients, that is,the compositions, identified in Table I on the X-axis. The Y-axisdepicts the data expressed as a percentage of tested IgE-R/MTT (“MTT” is3-(4,5-Dimethylthiazol-2yl)-2,5-Diphenyltetrazolium bromide) tountreated control cells IgE-R/MTT. The control IgE-R/MTT is acalculation to normalize IgE receptor expression to cellular viabilityusing well known and standard testing techniques.

As shown in FIG. 1, two compositions containing three ingredients, 3-4and 3-6 appeared very effective. The ingredients of those compositionsare Perilla seed, Picao preto, and Kiwi extract (3-4); and Perilla seed,Kiwi extract and Cinnamomum (3-6). It is surmised that the apparentincrease in the IgE receptor expression in cells treated with 4 and 5ingredients can be due to mild toxicity given the number of ingredients.

The methodology used to obtain the results of FIG. 1 will now bedescribed. First, RBL-2H3 cells, available from ATCC (American TypeCulture Collection) of Manassas, Va., were plated in 96 well plates at3×10⁴/well. Following adherence to the wells, the cells were exposed tothe sample combinations of ingredients listed in Table I, eachcombination referred to as a “sample.” Stock solutions of all sampleswere prepared in a solution of DMSO, ethanol, and water present in aratio of 5:3:2 at 100 mg/ml. Each sample was then diluted to 100 μg/mlin media. Duplicate plates of each sample were prepared so thatviability could be determined multiple times for each sample, with theresults of the multiple samples averaged following treatment of thecells.

The plates were incubated an additional 48 hours. To measureIgE-receptor expression, the cells were fixed for 2 hours with 1%gluteraldehyde. Following fixation, the cells were washed twice withPBS-Tween 20 (0.01%), available from Aldrich of Milwaukee, Wis. Proteinbinding sites were blocked for 2 hours with 2% non-fat dry milk in PBS(Phosphate Buffer Saline buffered at pH 7.3). The cells were washedtwice. The primary antibody, Rabbit anti-FcεRIα, available from UpstateGroup, Inc., Lake Placid, N.Y., was added to the wells and diluted1:4000 with 0.2% BSA (Bovine Serum Albumin). Following a 2 hourincubation, the wells were washed 5× with PBS-Tween 20 (0.01%)solutions. The secondary antibody, Goat anti-rabbit IgG-peroxidase,available from Sigma-Aldrich of St. Louis, Mo., was added to the wellsat 1:20000 with 0.2% BSA.

Following a 1 hour incubation the wells were washed five times. 100μliters (microliters) of TMB (3,3′,5,5±Tetramethyl benzidinedihydrochloride) Substrate, available from Chemicon International ofTemecula, Calif., was added to the wells. Subsequent color developmentwas stopped by the addition of a sufficient amount, H₂SO₄ (for example,100 μliters at 1 Normal).

For each of the multiple samples of the combination, the OD₄₅₀ (OpticalDensity measured at a wavelength of 450 nanometers) was read on a platereader.

Continuing with a description of the methodology to obtain the data inFIG. 1, the viability of the cells was next determined by incubating thecells in the second plate with MTT in a concentration of 1 mg/ml for 3hours. The reduced MTT was extracted with 70% isopropanol and the OD₅₄₀(Optical Density at a wavelength of 540 nanometers) was read for each ofthe multiple samples.

Again, the vertical bars for each sample in FIG. 1 represent the abilityof each sample, i.e., each composition, to control IgE receptorexpression on the RBL-2H3 cells relative to IgE receptor expression onuntreated RBL-2H3 cells. For example, the combination 3-4 exhibited anIgE receptor expression that was about 90% of the IgE receptorexpression of the untreated or control cells. In other words,combination 3-4 decreased IgE receptor expression by about 10% from whatthat expression was in untreated control cells.

It is believed that the results of the testing on the RBL-2H3 cells inthis example will be analogous to and/or indicative of the effect of thecompositions on IgE receptor expression that would occur on mast cellswithin the body of a mammal, such as a human. This belief is derivedfrom initial experiments, the results of which are illustrated in FIG.2. There, the X-axis represents the OD₄₅₀ values, and the Y-axisrepresents the expression of type I and type II IgE receptors at varyingdilutions. In particular, confirmation of IgE-RI, and not IgE-RII,expression on RBL-2H3 cells is shown in FIG. 2. Receptor expression wastested using the method described above with the rabbit anti-FcεRIIαantibody using antibody dilutions from 1:50 to 1:10000. The differentdilutions of the antibody were used to optimize the experimental method.A mouse monoclonal antibody against anti-FcεRIIα antibody, availablefrom Biodesign International of Saco, Me., was also used to test for thepresence of IgE-RII (RII) on the cells, and the data in FIG. 2 show thatlevels of the type II (RII) receptor are negligible, but that the levelsof type I receptor (RI) are significant. Because the type I receptor isthe receptor that mediates degranulation of mast cells, there is asignificant probability that the RBL-2H3 cells are appropriate forscreening for the down-regulatory effect on IgE receptor expression.Accordingly, it is believed that those tested compositions having theability to reduce IgE expression in the in-vitro testing above will besuitable for reducing IgE expression in animal subjects.

EXAMPLE 2

Multiple individual ingredients of the composition were tested todetermine the effectiveness of each ingredient at controlling IgEreceptor expression. Each of the individual ingredients in Tables II andIII below were tested using the methodology presented above inconnection with the combinations of ingredients in Example 1. The dataexpressed as Percent Control IgE Receptor/Viability denotes the level ofIgE receptor expression of treated cells relative to untreated controlcells in each experiment. The ingredients tested show different levelsof satisfactory reductions in IgE receptor expression. TABLE II Effectof Individual Ingredients on IgE Receptor Expression Percent Control IgEIngredient Receptor/Viability Perilla seed 81% Cinnamon 86% Picao preto91% Hesperidin 80% 90% Guaco 85% Holy Basil 87% Kakadu concentrate 83%Solarnum 85%

In addition to the ingredients in Table II, other ingredients that canbe used include Kiwi extract, Kiwi juice, and Acerola Cherry. The HolyBasil of Table II was obtained from GeniHerbs. TABLE III Effect ofIndividual Ingredients on IgE Receptor Expression Percent Control IgEIngredient Receptor/Viability Cumaru 91% Marigold 92% Nettle Root 91%Tumeric 89% Allergy Formula 83%

In addition to the ingredients listed in Table III, other ingredientsthat can be used include Ginger PE 5%, Boswin 30, Trikartu extract,BioPerine, as well as those ingredients in Example 5 below.

EXAMPLE 3

Multiple combinations of ingredients presented in Table II above weretested to determine the combinations effectiveness in controlling IgEsecretion by cells. FIG. 3 demonstrates the effects of the compositioncombinations on IgE secretion by cells relative to IgE secretion ofcells untreated with the compositions. Specifically, FIG. 3 presents thecombination of ingredients, that is, the compositions, identified inTable I on the X-axis. The Y-axis depicts the data expressed as apercentage of the levels of IgE secreted by control cells, that is cellsto which no compositions were added.

As shown in FIG. 3, the most effective compositions were 3-1, 3-4, 3-5,3-6 and 3-7. Those compositions comprise: Perilla seed, Hesperidin 80%,and Picao preto (3-1); Perilla seed, Picao preto, and Kiwi extract(3-4); Perilla seed, Picao preto, and Cinnamomum (3-5); Perilla seed,Kiwi extract, and Cinnamomum (3-6); Heperidin 80%, Picao preto andCinnamomum (3-7). As evident in FIG. 3, the compositions with 4 or 5ingredients also appear to inhibit IgE secretion. It is believed thatthis can be the result of mild toxicity due to multiple ingredients.

The methodology used to obtain the results of FIG. 3 will now bedescribed. First, U266 human myeloma cells, available from ATCC ofManassas, Va. were plated in 96 well plates at 4×10⁴/well. The cellswere exposed to the sample combinations of ingredients listed in TableI, each combination referred to as a “sample.” Stock solutions of allsamples were prepared in a solution of DMSO, ethanol, and water presentin a ratio of 5:3:2 at 100 mg/ml. Each sample was then diluted to 100μg/ml in media. Duplicate plates of each sample were prepared so thatviability could be determined multiple times for each sample, with theresults of the multiple samples averaged following treatment of thecells.

The plates were incubated an additional 48 hours. Thereafter, the amountof IgE in the supernatants was measured using a conventionalquantitative IgE ELISA kit available from Bethyl Laboratories ofMontgomery, Texas. Next, the measured OD450 data from the IgE ELISA wasinputted into a calibration graph, illustrated in FIG. 4, to determinethe concentration of IgE in ng/ml in each supernatant. This calibrationgraph was generated by measuring the OD450 of multiple, knownconcentrations (in nanograms per ml) of purified IgE. As shown in thegraph, the X-axis indicates the concentrations of the IgE in ng/ml; andthe Y-axis indicates the OD450 derived from each of the concentrations.The line 100 connects the data points, while line 110 is a trend linecorresponding to the equation in this particular embodiment of:Y=0.2909X−0.2527   Equation 1

With the IgE calibration graph, the amount of IgE for each sample wasdetermined, and the subsequent data was input into FIG. 3 forcomparative analysis.

It is believed that the effect of the compositions in Table 1 on IgEsecretion of U266 human myeloma cells is analogous to and indicative ofthe effect of those compositions on human or animal cells in-vivo due tothe similar cellular structure and allergic reaction of the test cellsand in-vivo cells.

EXAMPLE 4

Multiple individual ingredients of the composition were tested todetermine their ability to down regulate IgE receptor expression and thesecretion of IgE antibodies. These ingredients were Acerola cherryextract, Kiwi juice extract, Vitamin C and Holy Basil at varyingconcentrations. The Acerola extract and Vitamin C were obtained fromNutrilite of Lakeview, Calif.; and the Kiwi juice and Holy Basil wereobtained from P.L. Thomas & Co., Inc. of Morristown, N.J.

The data from this example demonstrate the effects of the differentingredients (samples) at varying concentrations on IgE receptorexpression as illustrated in FIG. 5. As shown there, Vitamin C had asignificant effect on IgE receptor expression. At 200 micrograms permilliliter, Vitamin C induced a 15% decrease in IgE receptor expression.The results illustrated in FIG. 5 were obtained using the samemethodology as set forth above in Example 1, except that theconcentration of each ingredient was varied as shown along the X-axis ofFIG. 5.

The data in FIG. 6 is representative of the effect of the variousingredients on inhibiting IgE secretion. The data are expressed aspercent control of IgE secreted by treated cells relative to untreatedcontrol cells. As demonstrated in FIG. 6, Acerola extract, Kiwi juice,Vitamin C and Holy Basil all inhibited IgE secretion in the amountsshown. Acerola extract was the most potent at inhibiting IgE secretion.The results illustrated in FIG. 6 were obtained using the samemethodology as set forth in Example 3, except that the concentration ofeach ingredient was varied as indicated on the X-axis of FIG. 6.

EXAMPLE 5

Multiple individual ingredients of the composition were tested todetermine the effectiveness of each ingredient at controlling IgEreceptor expression. Each of the individual ingredients in Table IVbelow were tested using the methodology presented above in connectionwith the combinations of ingredients in Example 1. The data expressed asPercent Control IgE-Receptor denotes the level of IgE receptorexpression of treated cells relative to untreated control cells in eachexperiment. The ingredients tested show different levels of satisfactoryreductions in IgE receptor expression. TABLE IV Effect of Ingredients onIgE Receptor Expression Ingredient Percent Control IngredientConcentration IgE-Receptor Luteolin 100 μg/ml 82 Cinnamomum 100 μg/ml 86Acerola 100 μg/ml 107 Tinospora 100 μg/ml 109

The data from Table IV is illustrated in FIG. 7 to graphicallydemonstrate the effects of the different ingredients. As shown there,luteolin from Perilla leaf and Cinnamon both had a significant effect onIgE receptor expression.

EXAMPLE 6

Multiple individual ingredients, as well as the Blend (which includesequal parts Cinnamon, Acerola and Picao) were tested to determine theeffectiveness of each at controlling IgE secretion using the methodologypresented above in connection with the ingredients in Example 3. Thedata in Table V expressed as Percent Control IgE Secretion denotes thelevel of IgE secretion by treated cells relative to untreated controlcells in each experiment. The ingredients tested show different levelsof satisfactory reductions in IgE secretion. TABLE V Effect ofIngredients on IgE Secretion Ingredient Percent Control IngredientConcentration IgE-Secretion Luteolin 100 μg/ml 53 Acerola 100 μg/ml 60Butterbur 100 μg/ml 71 Blend 100 μg/ml 73 Tinospora 100 μg/ml 77 Picao100 μg/ml 89 Cinnamon 100 μg/ml 94

The data from Table V is illustrated in FIG. 8 to graphicallydemonstrate the effects of the different ingredients. As shown there,most of the individual ingredients, as well as the Blend had asignificant effect on IgE secretion.

B. Allergy Mediator Testing and Results

The effects of the composition on allergy mediator release can beanalyzed in any in-vivo or in-vitro model. In an exemplary in-vitromodel, allergy mediator release was assessed following exposure of mastcells to the composition as well as individual ingredients of thecomposition. The cells were sensitized and stimulated to causedegranulation. After a predefined incubation period, the levels ofallergy mediators were measured with immunochemical methods. Thefollowing examples describe how mediator release was measured toidentify and select combinations of ingredients having acceptablemediator release inhibition. The methodologies set forth in thefollowing examples can be used to identify individual ingredients andcombinations of ingredients that have an effect on mediator release. Asan additional consideration, selection of individual ingredients andcombinations of ingredients for the composition can be based on theingredients' or combinations' ability to affect mediator release, aswell as the ability to down regulate IgE. Indeed, those ingredients orcombinations of ingredients that perform well at both mediator releaseand IgE down regulation are well suited for the treatment of allergicresponses.

EXAMPLE 7

Multiple ingredients and combinations of ingredients of the compositionpresented in Table VI were tested to determine the effectiveness of eachin inhibiting allergy mediator release. The mediators tested werehistamine, PGD₂ and LTC₄. The concentration of each ingredient orcombination for each mediator is also presented in the table. Forexample, the concentration of Rosmarinic acid when testing histamine was10 nanograms per milliliter, when testing PGD₂ was 1 microgram permilliliter, when testing LTC₄ was 0.1 micrograms per milliliter. TABLEVI Effect of Ingredients on Allergy Mediator Release Percent PercentPercent Ingredient Control of Ingredient Control Ingredient ControlConc. for Histamine Conc. for of PGD₂ Conc. For LTC₄ IngredientHistamine Release PGD₂ Release LTC₄ Release Rosmarinic  10 ng/ml 9.3  1μg/ml 94 0.1 μg/ml   120 acid Tinospora 100 ng/ml 10 10 μg/ml 94 1 μg/ml75 Blend 100 ng/ml 29 10 μg/ml 102 1 μg/ml 56 Luteolin 100 ng/ml 10 10μg/ml 68 1 μg/ml 157 Aframomum 100 ng/ml 24 10 μg/ml 68 1 μg/ml 88Acerola  1 μg/ml 70 0.1 μg/ml  70 10 μg/ml  63 Cinnamon  1 μg/ml 84 0.1μg/ml  84 10 μg/ml  60 Picao preto  1 μg/ml 96 N/A N/A N/A N/A

The data from Table VI is illustrated in FIG. 9 to graphicallydemonstrate the effects of the different ingredients on allergy mediatorrelease. As shown there, most of the individual ingredients, as well asthe Blend had a significant effect on mediator release. Moreover, anunexpected synergy was observed when certain ingredients were combined.For example, when Cinnamon, Acerola and Picao preto were combined toform the Blend, the Blend inhibited histamine release to 29% of thehistamine release that occurred in untreated cells. In contrast,Cinnamon individually inhibited histamine release to 84%, Acerola to 70%and Picao preto to 96%.

The methodology used to obtain the results of FIG. 9 will now bedescribed. First, RBL-2H3 cells, available from ATCC (American TypeCulture Collection) of Manassas, Va., were plated in 24 well plates at5×10⁴/well. Following adherence, the cells were exposed to theingredients or combination of ingredients noted in FIG. 9, also referredto as samples. The samples were used at the concentrations noted inTable VI. Stock solutions of all samples were prepared in a solution ofDMSO, ethanol, and water present in a ratio of 5:3:2 at 100 mg/ml. Eachsample was then diluted in media to the final concentration. The cellswere then incubated overnight with the samples. The following day amouse anti-dinitrophenyl (DNP) IgE antibody, available from Sigma of St.Louis, Mo., was added to the cells at a final concentration of 1 ng/ml.The cells were incubated for 1.5 hr at 37° C. to allow binding of theIgE molecule to the cellular IgE receptor via the constant region of theantibody.

Following this incubation period, the cells were washed twice. Asolution containing DNP conjugated to BSA (DNP-BSA), available fromCalbiochem of San Diego, Calif., was added to the cells at 0.5 ng/ml.The cells were incubated an additional 1 hr at 37° C. The supernatantswere assayed for the presence of histamine, PGD₂ and/or LTC₄ using ELISAkits commercially available from Cayman Chemical of Ann Arbor, Michiganaccording to the manufacturer's specifications.

Next, the measured OD450 data from the ELISA was inputted into acalibration graph to determine the amount by which histamine was reducedrelative to an untreated control sample. An example of a histaminecalibration graph is illustrated in FIG. 10. Suffice it to say thatcalibration graphs for PGD₂ and LTC₄ are similar. The calibration graphin FIG. 10 was generated by measuring the OD450 of multiple, knownconcentrations of purified histamine. As shown in the graph, the X-axisindicates the concentrations of the histamine in nanomolar; and theY-axis indicates the OD450 derived from each of the concentrations. Theline 120 connects the data points, while line 130 is a trend linecorresponding to the equation in this particular embodiment of:Y=−0.0025X ²+0.2915X+2.0775   Equation 2

With the histamine calibration graph, the amount of histamine for eachsample was determined, and the subsequent data was input into Table VIand FIG. 9 for comparative analysis.

It is believed that the effect of the compositions in this example, aswell as Examples 8 and 9, on allergy mediator release is analogous toand indicative of the effect of those compositions on human or animalcells in-vivo due to the similar cellular structure and allergicreaction of the test cells and in-vivo cells.

EXAMPLE 8

Multiple ingredients and combinations of ingredients of the compositionpresented in Table VII were tested to determine the effectiveness ofeach in inhibiting allergy mediator release. The mediator tested washistamine. The concentration of each ingredient or combination testedwas 10 nanograms per milliliter. For the Blend, the concentration of theindividual ingredients was equal, for example, Cinnamon was 3.33 ng/ml,of Acerola was 3.33 ng/ml and for Picao preto was 3.33 ng/ml. Nothingwas added to the control. Where ingredients were combined, each waspresent in an equal amount. For example, where the Blend was combinedwith Luteolin, both were present in equal amounts, that is, the Blendwas present in a concentration of 10 ng/ml, and Luteolin at 10 ng/ml.TABLE VII Effect of Ingredients on Histamine Release Percent Control ofIngredient Histamine Ingredient Concentration Release Rosemary 10 ng/ml76.7% Aframomum 10 ng/ml 95.6% Tinospora 10 ng/ml 95.0% Luteolin 10ng/ml 88.1% Butterbur 10 ng/ml 74.6% Control 10 ng/ml 100.0% Blend 10ng/ml 72.8% Blend + Rosemary 10 ng/ml 99.8% Blend + Butterbur 10 ng/ml105.7% Blend + Aframomum 10 ng/ml 99.9% Blend + Tinospora 10 ng/ml 78.0%Blend + Luteolin 10 ng/ml 58.4% Tinospora + Rosemary 10 ng/ml 82.4%Tinospora + Luteolin 10 ng/ml 72.2% Luteolin + Butterbur 10 ng/ml 75.3%Tinospora + Aframomum 10 ng/ml 88.3% Tinospora + Butterbur 10 ng/ml93.9% Luteolin + Aframomum 10 ng/ml 93.0% Butterbur + Aframomum 10 ng/ml73.8%

Select data from Table VII is illustrated in FIGS. 11-14 to graphicallydemonstrate the effects of the different ingredients on histaminerelease, as well as demonstrate the effect on histamine release whencertain ingredients are combined. The methodology used to produce theinformation of Table VII is identical to that in Example 7.

As shown in the Table and FIGS. 11-14, most of the individualingredients, as well as the Blend had a significant effect on mediatorrelease. Moreover, further unexpected synergy (in addition to thatobserved with the Blend) was observed when certain ingredients werecombined. For example, as shown in FIG. 11, the combination of the Blendwith luteolin inhibits histamine release to about 58% as compared to anuntreated control, whereas individually luteolin from Perilla leafinhibits to about 88%, and the Blend to about 73%. Further, thecombination of the Blend with tinospora inhibits histamine release toabout 78%, whereas individually the Blend inhibits histamine release toabout 73% and tinospora to about 95%.

Another example of unexpected synergy is shown in FIG. 12 via thecombination of tinospora with luteolin. There, the tinospora andluteolin combination inhibits histamine release to about 72%, whereasindividually tinospora inhibited histamine release to about 95%, andluteolin to about 88%.

Yet another example of unexpected synergy is shown in FIG. 13 via thecombination of the aframomum with butterbur. There, the combinationinhibits histamine release to about 74%, whereas individually aframomuminhibits histamine release to about 73% and butterbur to about 75%.

C. Combined IgE and Allergy Mediator Testing and Results

For certain ingredients and combination of ingredients of thecomposition, both IgE down regulation and allergy mediator release wereassessed contemporaneously. In an exemplary in-vitro model, both IgEdown regulation and allergy mediator release were assessed following themethodologies presented in Examples 1 and 7, respectively. The followingexamples identify the compositions tested, and present the results ofthe testing.

EXAMPLE 9

Multiple ingredients and combinations of ingredients of the compositionpresented in Table VIII were tested to determine the effectiveness ofeach in both down regulating IgE and inhibiting histamine release. Asused herein, the combination of luteolin (optionally from Perilla seed),Cinnamon and Acerola is referred to as the Formula. The concentration ofeach ingredient or combination of ingredients is noted in Table VIII.The concentration of each individual ingredient in the Formula wasequal. For example, the Formula IgE-R (IgE receptor expression)concentration was 100 μg/ml. Thus, the concentration of luteolin fromPerilla seed was 33.3 μg/ml, from Cinnamon was 33.3 μg/ml, and fromAcerola was 33.3 μg/ml. TABLE VIII Effect of Ingredients on IgE ReceptorExpression (IgE-R); IgE Secretion (IgE) and Histamine Release(Histamine) Ingredient Percent Ingredient Conc. Control Luteolin fromPerilla seed IgE-R 100 μg/ml 90.1% Cinnamon IgE-R 100 μg/ml 119.9%Acerola IgE-R 100 μg/ml 105.3% Formula IgE-R 100 μg/ml 89.0% Luteolinfrom Perilla seed IgE 100 μg/ml 95.5% Cinnamon IgE 100 μg/ml 100.2%Acerola IgE 100 μg/ml 77.9% Formula IgE 76.0% Luteolin from Perilla seed 1 μg/ml 99.9% Histamine Release Cinnamon Histamine Release  1 μg/ml77.5% Acerola Histamine Release  1 μg/ml 58.6% Formula Histamine Release 1 μg/ml 56.1%

The data from Table VIII is illustrated in FIG. 14 to graphicallydemonstrate the effects of the different ingredients on IgE receptorexpression, IgE secretion and histamine release. As shown in Table VIIIand FIG. 14, most of the individual ingredients exhibited a positiveeffect on IgE receptor expression, IgE secretion and histamine releasemediator release. However, surprising and unexpected synergy wasobserved when the ingredients were combined in the Formula.Specifically, the Formula inhibits histamine release as compared to acontrol (untreated with ingredients) to about 58%, whereas individuallyLuteolin inhibits histamine release to about 100%, Cinnamon to about 77%and Acerola to about 58%.

Further unexpected synergy is exhibited with respect to IgE downregulation. For example, the Formula down regulated IgE receptorexpression to about 89%, whereas individually Luteolin inhibits IgEreceptor expression to about 90%, Cinnamon to about 119% and Acerola toabout 105%. As a further example, the Blend down regulated IgE secretionto about 76%, whereas individually Luteolin inhibits IgE secretion toabout 95%, Cinnamon to about 100% and Acerola to about 78%.

The Formula may be combined with at least one of at least one of Picaopreto, Tinospora, Rosmarinic acid, and Aframomum to elicit similareffects on IgE receptor expression, IgE secretion and mediator release.

The above descriptions are those of the preferred embodiments of theinvention. Various alterations and changes can be made without departingfrom the spirit and broader aspects of the invention as defined in theappended claims, which are to be interpreted in accordance with theprinciples of patent law including the doctrine of equivalents. Anyreferences to claim elements in the singular, for example, using thearticles “a,” “an,” “the,” or “said,” is not to be construed as limitingthe element to the singular.

1. A composition comprising a Cinnamon ingredient, an Acerola ingredientand a Picao preto ingredient in an effective amount to inhibit anallergic response by at least one of down-regulating IgE expression,inhibiting IgE secretion and inhibiting an allergy mediator.
 2. Thecomposition of claim 1 comprising at least one of a Luteolin ingredient,an Aframomum ingredient, a Rosmarinic acid ingredient and a Tinosporaingredient.
 3. The composition of claim 2 comprising the Luteoliningredient.
 4. The composition of claim 2 comprising the Aframomumingredient.
 5. The composition of claim 2 comprising the Rosmarinic acidingredient.
 6. The composition of claim 2 comprising the Tinosporaingredient.
 7. The composition of claim 1 wherein the luteolin isderived from Perilla leaf.
 8. The composition of claim 5 wherein theRosmarinc acid is derived from Rosemary.
 9. The composition of claim 1wherein the Cinnamon ingredient, Acerola ingredient and Picao pretoingredient are present in an effective amount to inhibit the allergymediator, wherein the allergy mediator is at least one of prostaglandinD₂, Leukotriene C₄ and histamine.
 10. A composition comprising aCinnamon ingredient, an Acerola ingredient and a Luteolin ingredient inan effective amount to inhibit an allergic response by at least one ofdown-regulating IgE expression, inhibiting IgE secretion and inhibitingan allergy mediator.
 11. The composition of claim 10 comprising at leastone of a Picao preto ingredient, an Aframomum ingredient, a Rosmarinicacid ingredient and a Tinospora ingredient.
 12. The composition of claim11 comprising the Picao preto ingredient.
 13. The composition of claim11 comprising the Aframomum ingredient.
 14. The composition of claim 11comprising the Rosmarinic acid ingredient.
 15. The composition of claim11 comprising the Tinospora ingredient.
 16. The composition of claim 11wherein the Cinnamon ingredient, Acerola ingredient and Luteoliningredient are present in an effective amount to inhibit the allergymediator, wherein the allergy mediator is at least one of prostaglandinD₂, Leukotriene C₄ and histamine.
 17. The composition of claim 11wherein the Luteolin is derived from Perilla seed.
 18. A compositioncomprising a Luteolin ingredient and a Tinospora ingredient in aneffective amount to inhibit an allergic response by at least one ofdown-regulating IgE expression, inhibiting IgE secretion and inhibitingan allergy mediator.
 19. A method for inhibiting an allergic responsecomprising administering an effective amount of a composition includingat least three different ingredients selected from the group consistingof a Cinnamon ingredient, an Acerola ingredient, a Luteolin ingredientand a Picao preto ingredient, wherein the ingredients operate to atleast one of down-regulate IgE expression, inhibit IgE secretion andinhibit an allergy mediator.
 20. The method of claim 19 wherein theCinnamon ingredient, the Acerola ingredient, and Picao preto ingredientare present in the composition.
 21. The method of claim 19 wherein theCinnamon ingredient, the Acerola ingredient, and Luteolin ingredient arepresent in the composition.
 22. The method of claim 19 wherein thecomposition ingredients operate to inhibit an allergy mediator, and theallergy mediator is at least one of prostaglandin D₂, Leukotriene C₄ andhistamine.
 23. The method of claim 22 wherein the compositioningredients operate to inhibit histamine.